Ath 11 is complex: Raet1e and Esr1 as candidate genes; their mechanisms of action

Ath 11比较复杂：Raet1e和Esr1作为候选基因；

• 批准号: 8433465
• 负责人: JAN Leslie BRESLOW
• 金额: $ 50.17万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8433465
• 关键词: Affect; Aorta; Arterial Fatty Streak; Atherosclerosis; Biological Assay; Breeding; Candidate Disease Gene; Cause of Death; Cells; Cellular Stress; Chromosomes, Human, Pair 10; Complex; Coronary heart disease; Data; Developing Countries; Distal; ESR1 gene; Early treatment; Estrogen Receptor 1; Estrogens; FVB/N Mouse; Female; Gender; Gene Cluster; Gene Expression; Gene Expression Microarray Analysis; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Grant; Heritability; Histocompatibility Antigens Class I; Human; Immune; Immune system; Inbred Strain; Individual; Knockout Mice; Knowledge; Laboratories; Lesion; Ligands; MHC Class I Genes; Maps; Medical; Messenger RNA; Molecular; Mouse Strains; Mus; Myocardial Infarction; Nuclear Hormone Receptors; Pathway interactions; Physiological; Play; Population; Predisposition; Process; Proteins; Quantitative Trait Loci; Societies; Surface; Testing; Therapeutic; Transgenes; Transgenic Mice; Transgenic Organisms; Variant; Work; atherogenesis; atheroprotective; base; cell injury; congenic; cytokine; cytotoxicity; design; disability; genome wide association study; interest; macrophage; monocyte; mouse model; protein function; public health relevance; receptor; trait; transcription factor; tumor

DESCRIPTION (provided by applicant): Atherosclerosis is the major cause of death and disability in Westernized societies and is rapidly becoming so in developing nations. Despite significant medical progress our knowledge is incomplete and we still need to discover new genes and pathways in atherogenesis. Mouse models of atherosclerosis provide a complementary approach to human studies for such discoveries, and in an intercross between C57BL/6J and FVB/N mice on the apoEKO background we identified an atherosclerosis susceptibility locus on proximal mouse chromosome 10 (Ath11). Our work is at the forefront of the application of Quantitative Trait Locus (QTL) analysis not just to atherosclerosis but to all complex traits. We showed this locus contains 2 regions, a distal 10b region of 5 genes with Raet1e as a strong candidate gene, and a proximal 10a region of 21 genes with Esr1 an appealing candidate. The overall objective of this grant request is to confirm Raet1e and Esr1 as the culprit genes and elucidate their mechanisms of action. Raet1e is an MHC class I-like molecule expressed on the surface of stressed cells. It is a ligand for NKG2D, a receptor on the surface of NK, NKT, and T-Reg cells and some macrophages. This interaction plays a key role in the elimination of tumors and infected or damaged cells by the innate immune system. Our studies suggest aortic over expression of Raet1e is atheroprotective. To confirm this, we have made BAC Raet1e transgenic mouse strains and by breeding the transgene onto the apoEKO background we hope to prove our hypothesis. We will then identify the sequence variations between C57 and FVB in the Raet1e gene in the region of interest and find out which sequence variation is responsible. To determine how Raet1e over expression is atheroprotective we will analyze aortic gene expression patterns and cytokine profiles of mice with and without the Raet1e transgene. Using apoEKO and apoEKO NKG2DKO mice we will resolve if the effect of over expression of Raet1e on atherosclerosis is dependent or independent of the NKG2D receptor. We will identify by FACS analysis of splenic and aortic cells the immune cells involved in this process and with functional studies, such as cytotoxicity and proliferation assays, determine how over expression of Raet1e alters the physiological state of these cells. Esr1 is the main nuclear hormone receptor for estrogens. We will confirm Esr1 as the 10a region gene by expression analysis in subcongenic strains as well as comparing C57 and FVB Esr1 gene sequence, aortic mRNA and protein levels, and ESR1 function to identify the molecular and mechanistic basis of Esr1 as an atherosclerosis modifier gene. If we do not find support for Esr1 as a candidate gene for the 10a region we will identify, among the other 20 genes in the 10a region, the responsible gene by characterizing new subcongenic strains that further narrow the 10a region, and evaluate the most promising candidate genes by gene sequencing, gene expression analysis, and functional studies.

描述（由申请人提供）：动脉粥样硬化是西方社会死亡和残疾的主要原因，并且在发展中国家中迅速变得如此。尽管医学的进步很大，但我们的知识还是不完整的，我们仍然需要发现动脉粥样硬化中的新基因和途径。动脉粥样硬化的小鼠模型为这种发现提供了人类研究的补充方法，在Apoeko背景的C57BL/6J和FVB/N小鼠之间的互沿中，我们确定了近端小鼠染色体染色体10（ATH11）上的动脉粥样硬化易感性位置。我们的工作不仅是针对动脉粥样硬化的，而且对所有复杂性状的应用，都处于定量性状基因座（QTL）分析的最前沿。我们表明，该基因座包含2个区域，一个远端10b区域，由5个基因作为强候选基因，以及一个具有ESR1的2​​1个基因的近端10a区域，是一个吸引人的候选者。该赠款请求的总体目标是确认RAET1E和ESR1为罪魁祸首，并阐明其作用机制。 RAET1E是在应力细胞表面表达的MHC I类分子。它是NKG2D的配体，NKG2D是NK，NKT和T-REG细胞表面上的受体以及一些巨噬细胞。这种相互作用在消除肿瘤并被先天免疫系统感染或受损的细胞中起关键作用。我们的研究表明，主动脉对RAET1E表达的表达是动脉保护性的。为了证实这一点，我们已经制造了BAC RAET1E转基因小鼠菌株，并通过将转基因繁殖到Apoeko背景上，我们希望证明我们的假设。然后，我们将确定感兴趣区域中RAET1E基因中C57和FVB之间的序列变化，并找出哪些序列变化是造成的。为了确定在表达上的RAET1E是如何具有动脉保护性的，我们将分析有和没有RAET1E转基因的小鼠的主动脉基因表达模式和细胞因子谱。使用Apoeko和Apoeko NKG2DKO小鼠，如果Raet1e对动脉粥样硬化的过度表达对动脉粥样硬化的影响取决于或独立于NKG2D受体，我们将解决。我们将通过对脾细胞和主动脉细胞的FACS分析来识别参与此过程的免疫细胞以及功能研究（例如细胞毒性和增殖测定法），确定RAET1E的过度表达如何改变这些细胞的生理状态。 ESR1是雌激素的主要核激素受体。我们将通过在亚比例菌株中的表达分析以及比较C57和FVB ESR1基因序列，主动脉mRNA和蛋白质水平以及ESR1的功能来确定ESR1的分子和机械基础作为动脉粥样硬化基因基因。如果我们没有找到对10A区域的ESR1作为候选基因的支持，我们将通过表征10A个新的亚综合菌株的其他20个基因中的其他20个基因，从而进一步缩小10A区域，并通过基因测序，基因表达分析和功能研究来评估最有前途的候选基因。